Schwer B, Stunnenberg H G
European Molecular Biology Laboratory, Heidelberg, FRG.
EMBO J. 1988 Apr;7(4):1183-90. doi: 10.1002/j.1460-2075.1988.tb02929.x.
A cell free system mediating accurate transcription of vaccinia virus genes was established using lysates of cells in the late phase of infection. Vaccinia late genes are faithfully transcribed in this extract whereas cellular pol II and pol III promoters are not recognized. The late viral transcripts contain a poly(A) head of approximately 35 nt at the 5' end which is not co-linearly encoded in the externally added template. The transcripts obtained in vitro are indistinguishable from the mature in vivo RNAs. The poly(A) head is synthesized de novo and its formation appears to be directly coupled to the transcription of the gene. The synthesis of the poly(A) head transcripts in vitro is consistent with a proposed slippage model.
利用感染后期细胞的裂解物建立了介导痘苗病毒基因精确转录的无细胞系统。痘苗晚期基因在该提取物中能被忠实地转录,而细胞聚合酶II和聚合酶III启动子则不被识别。晚期病毒转录本在5'端含有一个约35个核苷酸的多聚(A)头部,该头部在外部添加的模板中并非共线性编码。体外获得的转录本与体内成熟RNA无法区分。多聚(A)头部是重新合成的,其形成似乎与基因转录直接相关。体外多聚(A)头部转录本的合成与提出的滑动模型一致。